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Search Completed | Title | Sensors and Actuators A 126 (2006) 455–462 Read-out concepts for multiplexed bead-based fluorescence immunoassays on centrifugal microfluidic platforms
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Text | Sensors and Actuators A 126 (2006) 455–462 Read-out concepts for multiplexed bead-based fluorescence immunoassays on centrifugal microfluidic platforms | 001
Sensors and Actuators A 126 (2006) 455–462
Read-out concepts for multiplexed bead-based fluorescence immunoassays on centrifugal microfluidic platforms
L. Riegger a,∗, M. Grumann a, T. Nann b, J. Riegler b, O. Ehlert b, W. Bessler c, K. Mittenbuehler c, G. Urban b,d, L. Pastewka a, T. Brenner a,
R. Zengerle a,e, J. Ducre ́e a,e
a IMTEK-University of Freiburg, Lab for MEMS Applications, Georges-Koehler-Allee 106, 79108 Freiburg, Germany b University of Freiburg, FMF, Germany
c University Hospital Freiburg, Institute for Molecular Medicine and Cell Research, Germany
d IMTEK-University of Freiburg, Lab for Sensors, Germany
e HSG-IMIT, Institute for Micromachining and Information Technology, Germany
Received 9 June 2005; received in revised form 31 October 2005; accepted 3 November 2005 Available online 28 December 2005
We present novel concepts to process and read out multiplexed, bead-based fluorescence immunoassays. At the start of the read-out process, a statistically arranged monolayer of color-encoded beads is aggregated in a detection chamber. Each bead is first identified by incorporated color tags which are either dyes or luminescing quantum dots (QDs). Subsequently, the reaction-specific fluorescence signal is quantified. The read-out process is accelerated by an in-house-developed image-processing algorithm. The optical read-out device consists of standard components, e.g. a color CCD-camera as detection unit, an LED as light source, optical filters, and a drive to spin the polymer disk. The liquid handling along the complete assay protocol is realized on a centrifugal lab-on-a-disk platform. We successfully demonstrate the performance of this device by the implementation of a hepatitis A and a tetanus assay.
© 2005 Elsevier B.V. All rights reserved.
Keywords: FIA; Multiplexed; Beads; Quantum dots; Centrifugal microfluidics; Rotation
The strong trend towards decentralized point-of-care tech- nologies in medical diagnostics has stimulated the development of miniaturized “lab-on-a-chip” systems [1–4]. These labs-on- a-chip feature a set of basic unit operations such as sample injection, separation, metering, mixing, and reacting to integrate and thus automate full diagnostic test protocols on a credit card- sized microfluidic substrate. Many complex tasks have already been realized, among them simultaneous DNA amplification and detection , HIV immunoassays , fully integrated single-cell processing , and combinatorial chemistry . Apart from the process integration, the major benefits of lab-on-a-chip tech- nologies are the minute consumption of sample and reagents,
∗ Correspondingauthor.Tel.:+497612037312;fax:+497612037322. E-mail address: email@example.com (L. Riegger).
0924-4247/$ – see front matter © 2005 Elsevier B.V. All rights reserved. doi:10.1016/j.sna.2005.11.006
short times-to-result, as well as their amenability for multiplex- ing and parallelization.
We here consider “lab-on-a-disk” technologies which utilize centrifugal forces for the transport of fluids. These “labs-on-a- disk” have been investigated by many groups [9–16], and several commercial products have been launched to the market [17–20]. We here for the first time investigate multiplexed immunoassays carried out on a microfluidic “lab-on-a-disk.” Our modular cen- trifugal platform [21,22] is constituted by a passive microstruc- tured disk as a disposable part and by a reusable centrifuge and detection unit. Our novel read-out technique enables a multi- plexed screening of a set of fluorescence immunoassays within a single channel and for several channels in parallel.
This paper is structured in the following way. We first describe the principle of a multiplexed assay. Next, all experimental components are outlined, followed by a discussion of the imple- mented read-out strategies. Finally, we present the results of two immunoassays and conclude.
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